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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
Statistical optimization of a podoviral anti-MRSA phage CCASU-L10 generated from an under sampled repository: Chicken
Israa M Abd-Allah1, Ghadir S El-Housseiny1, Mohamed H Al-Agamy2,3
1Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University, Organization of African Unity Street, Abbassia, Cairo, Egypt.
Introduction:
The insurgence of antimicrobial resistance is an imminent health danger globally. A wide range of challenging diseases are attributed to methicillin-resistant Staphylococcus aureus (MRSA) as it is weaponized with a unique array of virulence factors, and most importantly, the resistance it develops to most of the antibiotics used clinically. On that account, the present study targeted the optimization of the production of a bacteriophage active against MRSA, and evaluating some of its characters.
Methods And Results:
The bacteriophage originated from a quite peculiar environmental source, raw chicken rinse and was suggested to belong to Podoviridae, order Caudovirales. It withstood a variety of extreme conditions and yield optimization was accomplished via the D-optimal design by response surface methodology (RSM). A reduced quadratic model was generated, and the ideal production conditions recommended were pH 8, glycerol 0.9% v/v, peptone 0.08% w/v, and 107 CFU/ml as the host inoculum size. These conditions led to a two-log fold increase in the phage titer (1.17x10¹² PFU/ml), as compared to the regular conditions.
Discussion:
To conclude, statistical optimization successfully enhanced the output of the podoviral phage titer by two-log fold and therefore, can be regarded as a potential scale-up strategy. The produced phage was able to tolerate extreme environmental condition making it suitable for topical pharmaceutical preparations. Further preclinical and clinical studies are required to ensure its suitability for use in human.
Insights
This study optimized bacteriophage production against methicillin-resistant Staphylococcus aureus (MRSA), achieving a two-log fold increase in titer. The enhanced phage is stable in extreme conditions, showing potential for topical pharmaceutical use.
Area of Science:
- Microbiology
- Virology
- Antimicrobial Resistance
Background:
- Antimicrobial resistance, particularly from methicillin-resistant Staphylococcus aureus (MRSA), poses a significant global health threat.
- MRSA infections are challenging due to virulence factors and antibiotic resistance.
- Bacteriophages offer a potential alternative to antibiotics for combating resistant bacteria.
Purpose of the Study:
- To optimize the production of a bacteriophage effective against MRSA.
- To evaluate the characteristics of the produced bacteriophage.
- To explore bacteriophage therapy as a strategy against MRSA.
Main Methods:
- Isolation of a bacteriophage from raw chicken rinse, tentatively identified as Podoviridae, order Caudovirales.
- Optimization of bacteriophage production using response surface methodology (RSM) with a D-optimal design.
- Determination of optimal production conditions: pH 8, 0.9% glycerol, 0.08% peptone, and 10^7 CFU/ml host inoculum.
Main Results:
- The optimized conditions resulted in a two-log fold increase in bacteriophage titer, reaching 1.17x10^12 PFU/ml.
- The bacteriophage demonstrated stability under various extreme environmental conditions.
- The phage production was successfully scaled up using statistical optimization.
Conclusions:
- Statistical optimization significantly enhanced bacteriophage yield, presenting a viable scale-up strategy.
- The phage's tolerance to extreme conditions suggests suitability for topical pharmaceutical applications.
- Further preclinical and clinical studies are necessary to validate its therapeutic potential in humans.
